JPS6143981A - Continuous flow homogenizer - Google Patents

Abstract

PURPOSE:To obtain the titled homogenizer having high disintegration efficiency, and operble easily, by placing a pestle rotatably in a tubular vessel having openings at the upper and the lower ends, and jointing the rotary shaft of the pestle to the driving shaft with a coupling. CONSTITUTION:A transparent tubular vessel 9 having the inlet 5 of unemulsified tissue at the tip and the outlet 7 of the emulsified tissue at the bottom, is attached detachably to the supporting rod 3, and a pestle 13 is placed in the vessel interposing a proper small gap between the inner wall of the vessel 9. The rotary shaft 11 of the pestle 13 is jointed detachably to the driving shaft 17 with the coupling 19 to facilitate the attaching and detaching of the tubular vessel. The emulsifying operation can be carried out continuously by this apparatus.

Description

[Detailed Description of the Invention] [Industrial Application Field] The present invention relates to a homogenizer used to disrupt cell membranes and separate intracellular organelles, which has a simple configuration, can be manufactured at low cost, and has high disruption efficiency. , relates to a continuous flow homogenizer that is convenient to handle.

[Description of Prior Art] In biology, medicine, and other research fields, it is necessary to separate intracellular Golgi bodies, organelles such as mitochondria, and other enzymes. To do this, cells must be disrupted by some means, but conventional cell membrane disruption devices use ultrasonic waves, apply pressure with high-pressure nitrogen gas, etc., and rapidly remove this pressure. There are also methods that crush cell membranes by applying pressure to the cell fluid using a hydraulic machine and passing it through pores, called the Botta type. Methods of dividing cells by applying motion, and methods of rotating a pestle inside a tubular container using an existing rotary machine have also been used.

[Problems with the Prior Art] These conventional cell membrane disruption devices generally have poor disruption efficiency and are inconvenient to handle. For example, in the widely popular pot type, the tubular container must be moved up and down manually, which is inefficient.

For example, other devices using ultrasonic waves or high-pressure nitrogen require large and expensive devices.

[Means for Solving the Problems] The present invention solves the problems of the prior art described above, and provides a homogenizer for cell membrane disruption that has a simple device configuration, can be manufactured at low cost, has high disruption efficiency, and is convenient to handle. The homogenizer is characterized by having a tubular container having an unemulsified tissue inlet at the upper end and an emulsified tissue outlet at the lower end removably supported on a support. A pestle equipped with a rotating shaft is rotatably disposed inside the container with an appropriate gap, and a pestle is placed at the upper end of the tubular container by receiving a rotational driving force from a rotational driving section provided at the upper end of the column. A rotary drive shaft is provided vertically facing the pestle, and the rotary drive shaft and the rotary shaft provided on the pestle are removably connected to each other via a coupling.

[Operation] When using the homogenizer for disrupting cell membranes having the above configuration, first, for example, mince an animal sample material,
A buffer solution is added to obtain a cell solution, and the cell solution is poured into an unemulsified tissue inlet at the upper end of a tubular container supported by a column and equipped with a rotatable pestle. is rotated via a rotating drive shaft.

Then, the cell fluid is emulsified between the tubular container and the pestle, the cell membranes are crushed, and the cell fluid is sequentially discharged from the bottom outlet of the tubular container. The work of attaching or removing the tubular body container supported on the support l is carried out using the rotary shaft provided on the pestle,
The rotary shaft is removably connected to the rotating shaft via a coupling, so by removing this coupling, the tubular container can be removed without completely removing the pestle from the tubular container. It can be easily attached and detached.

In addition, since an unemulsified tissue inlet is provided at the upper end of the tubular container, unemulsified tissue fluid can be continuously introduced through this inlet and a continuous emulsification operation can be performed, making the emulsification work extremely efficient. can do well.

[Description of Embodiment] Hereinafter, one embodiment of the present invention will be described in detail using the drawings.

FIG. 1 is a perspective view of the entire continuous flow homogenizer, FIG. 2 is a sectional view of a tubular container equipped with a pestle, and FIG. 3 is a detailed view of the coupling.

In FIG. 1, a transparent tubular container 9 having an unemulsified tissue inlet 5 at the upper end and an emulsified tissue discharge lower at the lower end is removably supported on a support 3 erected on a base 1. A pestle 13 having a rotating shaft 11 is rotatably disposed inside the tubular container 9 with an appropriate gap between it and the inner wall of the tubular container 9, and a pestle 13 is provided at the upper end of the support 3. A rotary drive shaft 17 is provided which receives a rotary drive force from a rotary drive unit 15 and is suspended vertically toward the upper end of the tubular body container 9, and the rotary drive shaft 17 and the rotary shaft 11 provided on the pestle 13 are connected to each other. A continuous flow homogenizer is shown removably connected via a coupling 19.

For the base 1 and the support column 3, for example, a commercially available experimental stand may be used as is, but here, a rotation drive unit 15 is provided above the support column 3.

21 is an outlet, and 23 is a voltage exchange section.

Also, a mounting bracket 25 for holding the tubular body container 9 on the support column 3.
There are 27.

The mounting brackets 25 and 27 include semicircular arc-shaped fixing members 25a and 27a that open toward the tubular body container 9, and thumbscrews 25b and 2.
It consists of semicircular arc-shaped opening/closing members 25c and 27c symmetrical to the fixing member which is rotatably equipped with the fixing member 25.
A, 27a is provided with a tap hole into which the threaded portion of the thumb screw 25b, 27b is screwed, and the opening/closing member 2 is provided at the other end.
A hinge (not shown) is provided for rotating the fixing members 5b and 27b relative to the fixing members 25a and 27a. Butterfly screw 25b12
7b may be a so-called baratin tablet. Fixed member 25
A cushion (not shown) made of rubber or the like is provided inside the arcuate portions of the opening/closing members 25c, 27a and the opening/closing members 25c, 27c, respectively, so that the tubular container can be fixed flexibly, firmly, and reliably.

Note that if this cushion is made white or gray, the emulsified state can be observed through the transparent tubular container.
It is convenient because you can

On the front of the rotary drive unit 15, there is a switch 2 for power supply A/A.
9, a start switch 31, a stop switch 32, a digital switch 33 for setting the rotational speed, an LED (light emitting die board) type speed display device 135 for displaying the actual rotational speed, etc. are provided.

The rotary drive shaft 17 is rotated in one direction at a speed determined by the digital switch 33 via a transmission mechanism (not shown).

As shown in FIG. 2, an unemulsified tissue (cell fluid) inlet 5 is formed at the upper end of the tubular body container 9 and has an inverted conical opening to receive a large amount of tissue solution. An emulsified tissue discharge lower is provided at the bottom of the tubular body container 9.

An example of the tubular body container 9 made of glass is shown;
It is not limited to this.

FIG. 2 shows a state in which the pestle 13 is positioned approximately in the middle of the tubular body container 9, and in this state, a first chamber 35 containing unemulsified tissue (cell fluid) is located above the tubular body container 9. A second chamber 37 is formed below to guide the emulsified liquid to the discharge lower. The gap between the pestle 13 and the tubular container 9 is the distance d(0) necessary to separate the cell nucleus and other organelles.
, 0.05 to about 0.211), and is determined depending on the cell type.

In FIG. 2, the rotating shaft 11 provided on the pestle 13 is indicated by the arrow 3.
Rotate in direction 9. The pestle 13 is made of Teflon, for example, and the above-mentioned first to 35
A large number of guide grooves 41 and 41 are provided in the central part 13a of the pestle to allow the unemulsified tissue to be fed while gradually decreasing the distance from the inner circumferential surface of the tubular body container 9. Guide groove 41.4
The depth of the pestle is approximately 3-3 at the top, and gradually becomes shallower toward the center of the pestle until it reaches zero.

Since the pestle 13 is provided with the guide grooves 41, 41 as described above, as the pestle 13 rotates in the direction 39, the first chamber 3
The unemulsified tissue of No. 5 is the lower part of the pestle 13 and the tissue crushing part 13a.
will be sent towards. The cells trapped between the surface of the tissue disruptor m13d and the inner surface of the tubular body container 9 are ruptured by causing a gentle shearing action within a small gap.

A detailed view of the coupling 19 is shown in FIG.

The figure (a) is a perspective view showing an assembled state, and the figure (il) is an exploded perspective view thereof.

The coupling 19 includes an upper joint 43, an elastic joint 45,
The upper joint 43 consists of a lower joint 47 and a rotary drive shaft 1.
7 and the lower joint 47 are connected to the rotating shaft 11 with screws 49, respectively.
．． It is fixed using 49.

The upper joint 43 has small holes on both sides thereof, and the end of the clip 53, which has a holding part 51 that tilts downward and holds the lower end surface of the lower joint, is fitted into the small hole so that it can rotate freely. I support it. Further, below the upper joint 43 and above the lower joint, there are provided joint portions 57.57 of a height (for example, 511IIl) each provided with a detent gear 55.55.

The elastic joint 45 is made of rubber (rubber hardness 70 to 801-1s),
Alternatively, it is made of an elastic material such as plastic, and has a concave groove 5 therein that fits with each detent gear 55 of the joint part 57 provided in the upper joint 43 and the lower joint 47, respectively.
9.59 on both the upper and lower ends.

The elastic joint 45 may be fixed to either the upper joint 43 or the lower joint 47.

The drive shaft 17 is inserted into the upper joint 43 and fixed with screws 49, while the rotating shaft 11 is inserted into the lower joint 47 and fixed with screws 49. Therefore, by butting the upper joint 43 and the lower joint 47 from both the upper and lower end directions of the elastic joint 45, and lowering the clip 53 downward, the state shown in Fig. <a> is obtained. In this state, the rotating shaft 11 is connected to the driving shaft 17.
However, a deviation of a predetermined angle (approximately 3 degrees) is allowed.

Now, it is assumed that the emulsification work is completed and the tubular body container 9 is removed from the assembled state shown in FIG. 1. In this case, the clip 53 shown in FIG. 3 is rotated to remove the holding part 51 from the lower joint 47, and the pestle 13 integrated with the rotating shaft 11 is moved downward by the dimension shown in FIG. 3(b). lower to After that, the thumbscrew 25 of the mounting member 25.27
1), 27b may be loosened, the opening/closing member may be removed from the fixing members 25a, 250, and the tubular body container 9 may be removed.

When attaching the tubular body container 9 at the start of the emulsification operation, this operation may be performed in reverse. First, the tubular body container 9 is loosely attached to the mounting fitting 25. After that, the tubular container 9 is pushed down and positioned until the narrow diameter part (neck part) of the inverted cone of the unemulsified tissue inlet 35 (see Figure 2) of the tubular container 9 comes into contact with the upper end of the upper mounting bracket. do. Then thumbscrew 25
b, 27b are securely tightened, and the rotary shaft 11 attached to the pestle and the rotary drive shaft 17 are joined via the coupling 19 and fixed with the clip 53.

In this example, when attaching and detaching the tubular body container 9, the pestle 13 can be lowered or raised by the above-mentioned dimension, but if the detachable coupling 19 is not provided, the tubular body container 9 The container 9 must be removed from the pestle 13 in a fixed position, which is inconvenient. However, in this example, since the coupling 19 is provided as described above, the distance between the bottom surface of the base and the rotary drive unit 15 is set to the desired height L, thereby eliminating unnecessary height for removing the tubular body container 9. -ing

In addition to this example, it is conceivable that what is called a chuck may be used as another means for eliminating the need for the height at which the tubular body container is removed.

However, the chuck itself has no means for dealing with vibrations between the rotating shaft and the drive shaft, and although this chuck cannot be used in place of a coupling, it is of course possible to use these chucks in conjunction with a coupling.

[Effects of the Invention] The present invention provides a tubular container having an unemulsified tissue inlet at the upper end and an emulsified tissue outlet at the lower end and is detachably supported on a support, and has a rotating shaft inside the tubular container. A pestle equipped with a pestle is rotatably arranged with an appropriate gap, and a rotary pestle is suspended vertically toward the upper end of the tubular container by receiving a rotational driving force from a rotational driving part provided at the upper end of the pillar. The continuous flow homogenizer is characterized in that a drive shaft is provided, and the rotary drive shaft and the rotary shaft provided on the pestle are removably connected via a coupling.

Therefore, this apparatus has a simple structure and can be manufactured at low cost, and can perform emulsification work continuously and efficiently to obtain a high-quality emulsion. In addition, when you remove the device to clean, replace or repair the container or pestle, such as when moving on to the next task, you can easily remove the mounting bracket of the container and disengage the clip of the coupling. This has the advantage that the container and pestle can be taken out.

Note that the present invention is not limited to the embodiments described above, and it is of course possible to implement the present invention in aspects other than the embodiments described above by making appropriate design changes.

[Brief explanation of the drawing]

The drawings all show examples, and FIG. 1 is a perspective view of the entire continuous flow homogenizer device. FIG. 2 is an explanatory cross-sectional view of the tubular container with a pestle attached. FIG. 3 shows explanatory diagrams of the coupling, where (a) is a perspective view showing an assembled state, and (b) is an exploded perspective view. 1... Base 3... Support 5... Unemulsified tissue inlet a... Emulsified tissue outlet 9... Tubular body container 11.
... Rotation shaft 13 ... Pestle 15 ... Rotation drive unit 17 ... Rotation drive shaft 19 ... Coupling ceramics 6 exhibition f Figure 1 Figure 2 Figure 3 (a) Figure 3 (b ) ■

Claims (1)

[Claims] A tubular container having an unemulsified tissue inlet at the upper end and an emulsified tissue outlet at the lower end is removably supported on a post, and a pestle equipped with a rotating shaft is inserted into the interior of the tubular container. The rotary drive shaft is arranged to be freely rotatable with a gap therebetween, and receives a rotary drive force from a rotary drive unit provided at the upper end of the column, and is suspended vertically toward the upper end of the tubular body container. A continuous flow homogenizer characterized in that a shaft and a rotating shaft provided on the pestle are removably connected via a coupling.